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Related Concept Videos

DNA-only Transposons02:57

DNA-only Transposons

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
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Transposons01:24

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Transposons, or "jumping genes," are small mobile genetic elements (MGEs) that range from 700 to 40,000 base pairs in length. They are found in all organisms and can move within the same chromosome or transfer to different chromosomes. In some cases, transposons can also jump between different host DNA molecules, such as plasmids or viruses, contributing to genetic variability.Barbara McClintock first discovered these mobile genetic elements in the 1940s while studying maize genetics, and she...
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Overview of Transposition and Recombination02:13

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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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LTR Retrotransposons03:08

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LTR retrotransposons are class I transposable elements with long terminal repeats flanking an internal coding region. These elements are less abundant in mammals compared to other class I transposable elements. About 8 percent of human genomic DNA comprises LTR retrotransposons. Some of the common examples of LTR retrotransposons are Ty elements in yeast and Copia elements in Drosophila.
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Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis
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Transposon for protein engineering.

Vandan Shah1, Jin Ryoun Kim1

  • 1Othmer-Jacobs Department of Chemical and Biomolecular Engineering, New York University , Brooklyn, NY, USA.

Mobile Genetic Elements
|January 17, 2017
PubMed
Summary

Engineered Mu transposons offer an efficient method for protein engineering. These tools improve the construction of random insertional fusion and circular permutation libraries, overcoming limitations of conventional techniques.

Keywords:
MuRCP transposonMuST transposoncombinatorial libraryprotein engineeringrandom circular permutationrandom insertional fusion

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Area of Science:

  • Protein engineering
  • Molecular biology
  • Biotechnology

Background:

  • Protein insertional fusion and circular permutation are key protein engineering strategies.
  • Rational design for these techniques can be challenging, especially without structural data.
  • Existing combinatorial methods have drawbacks, including suboptimal linkers and sequence modifications.

Purpose of the Study:

  • To present engineered Mu transposons as a facile and efficient tool for protein engineering.
  • To overcome limitations of conventional methods for constructing random insertional fusion and circular permutation libraries.
  • To enable the creation of high-quality protein libraries.

Main Methods:

  • Development and application of engineered Mu transposons.
  • Construction of random insertional fusion libraries.
  • Construction of random circular permutation libraries.

Main Results:

  • Engineered Mu transposons alleviate limitations of conventional library construction methods.
  • Facilitates efficient creation of high-quality libraries for protein engineering.
  • Overcomes issues with inter-domain linker length, sequence modification, and ligation efficiency.

Conclusions:

  • Engineered Mu transposons provide a superior approach for protein engineering library construction.
  • This method enhances the creation of diverse protein functionalities and sequences.
  • Offers a significant advancement over traditional techniques for insertional fusion and circular permutation.